Functional Metagenomics to Mine Soil Microbiome for Novel Cadmium Resistance Genetic Determinants

Metal resistance genes are valuable resources for genetic engineering of bioremediation tools. In this study, novel genetic determinants involved in cadmium (Cd) resistance were identified using a small-insert metagenomic DNA library constructed from an arable soil microbiome. A total of 16 recombinant plasmids harboring 49 putative open reading frames (ORFs) were found to be associated with enhanced Cd tolerance. In addition to several ORFs for ion transport/chelation and stress response, most ORFs were assumed to be associated with non-direct metal resistance mechanisms such as energy metabolism, protein/amino acid metabolism, carbohydrate/fatty acid metabolism, and signal transduction. Furthermore, 13 ORFs from five clones selected at random were cloned and subject to Cd resistance assay. Eight of these ORFs were positive for Cd resistance when expressed in Escherichia coli, among which four ORFs significantly reduced Cd accumulation and one increased Cd enrichment of the host cells. Notably, C1-ORF1, potentially encoding a histidine kinase-like adenosine triphosphatase, was the most effective Cd resistance determinant and reduced host Cd accumulation by 33.9%. These findings highlight the vast capacity of soil microbiome as a source of gene pool for bioengineering. The novel genetic determinants for Cd resistance identified in this study merit further systematic explorations into their molecular mechanisms.     

Isolation, identification, and characterization of an aluminum-tolerant bacterium Burkholderia sp. SB1 from an acidic red soil

Aluminum (Al) toxicity usually occurs in acidic soils worldwide, which is detrimental to the growth of organisms. An Al-tolerant bacterium, SB1, was isolated from an acidic red soil of Chingkang Mountain, located in Jiangxi Province of China. Polyphasic analysis, including a 16S rDNA phylogenetic tree as well as morphological and physicochemical properties, revealed that the isolate was a gramnegative, rod-shaped bacterium, which was recognized as Burkholderia sp. SB1 and had extreme acidity tolerance (pH 2.2) and excellent Al resistance (270 mg L^-1 Al³⁺). It could remove Al by up to 97.7% at a concentration of 54 mg L^-1 Al³⁺. SB1 behavior under different temperatures and antibiotics was also examined. SB1 preferred moderate temperature conditions, ranging from 25 to 37℃, and exhibited notable resistance to multiple antibiotics (including ampicillin, streptomycin, and tetracycline), except for being sensitive to chloramphenicol. Therefore, as the first reported bacterium to possess favorable Al resistance and excellent Al removal, Burkholderia sp. SB1 can potentially be used as an agent for bioremediation of Al-contaminated acidic red soils.     

Enhanced Lead Uptake by an Association of Plant and Earthworm Bioaugmented with Bacteria

Lead (Pb) is recorded as the second most hazardous pollutant of the environment. Previous cases of Pb bioremediation has been reported using single biosystem, but very few reports are available in biological approaches using multi-biosystems to achieve an enhanced bioremoval of Pb. The present study evaluated the capacity of a unique association of Pennisetum purpureum, a hyperaccumulator plant, and Lumbricus terrestris (earthworm) bioaugmented with a Pb-resistant bacterium, obtained from an industrially contaminated site and identified as isolate VITMVCJ1 Klebsiella variicola, to bioremediate Pb. The Pb-resistant gene was amplified in the bacterial isolate VITMVCJ1. The study was conducted for 60 d. Results verified that the bioaugmentation process enhanced 1) root and shoot length of the plants, 2) chlorophyll content of the plants, and 3) biofilm-producing ability of the microbes from the rhizosphere region of the plants. The total phenolic and flavonoid contents were found to be lower in the plants in the bioaugmented setup. The study also observed a reduction in the toxic effects of Pb on earthworm and plant. The earthworm was used to assess the Pb-induced stress syndrome after exposure to sublethal concentrations of Pb in the soil. A reduction in the content of malondialdehyde, a potential biomarker, on exposure to Pb demonstrated the role of the inoculum to alleviate heavy metal-induced stress in earthworms. All three symbionts accumulated Pb; Pb was accumulated mainly in the root of the plant, and poorly in the shoot of the plant and body mass of the earthworm. The bioaugmentation system exhibited stable and excellent uptake of Pb from the contaminated soils. The results of the present study suggest the positive effect of the synergistic association of the plant and earthworm with appropriate microbes for the bioremoval of Pb.     

Preliminary assessment of large‐scale co‐culture of sandfish (Holothuria scabra) with the Babylon snail (Babylonia areolata) in earthen ponds and in raceways

Holothuria scabra (sandfish) and Babylonia areolata were trialed in two large‐scale co‐culture experiments. Experiment 1 assessed co‐culture in 4 × 400 m² earthen ponds where Babylonia were cultured in a central pen at a density of 400 individuals/m² with sandfish occupying the remaining pond space at 1.1 individuals/m². Sandfish grew from 18.20 ± 6.67 g to 119.03 ± 17.74 g in 92 days and Babylonia (fed trash fish in both experiments) grew from 0.90 ± 0.38 g to 4.93 ± 1.44 g, but Babylonia growth was not increased in co‐culture compared to monoculture. Water and sediment quality varied between co‐culture and monoculture ponds. Neither showed clear improvement due to sandfish culture. Experiment 2 compared non‐segregated sandfish‐Babylonia co‐culture with Babylonia monoculture in 20 m² concrete raceways. Sandfish were cultured at 2 individuals/m² and Babylonia at 300 individuals/m². Sandfish grew up to 1.91 g.day^?1 with 100% survival. Babylonia weight gain was significantly greater in co‐culture raceways (3.35 ± 0.64 g over 61 days), which was double that of Babylonia in monoculture. Substrate total N was reduced by 20% in co‐culture compared with monoculture (p = .032). This provisional study of commercial scale sandfish‐Babylonia co‐culture demonstrates culture compatibility, providing a basis for further system development.     

不同工业大麻品种对田间5种重金属吸收积累特性的比较

为了分析不同工业大麻品种对不同重金属富集与转移的差异，以云南5个工业大麻主栽品种（ym1~ym5）为试验材料，修复云南矿区重金属污染严重的农田。结果表明： 5个工业大麻品种中，成熟期的根系对Pb和Cd吸收量最大的为ym1，对As、Cu和Zn吸收量最大的为ym3，而ym5对Pb、Cu、Cd和Zn吸收量皆最小；茎叶对As、Cu和Cd吸收量最大的为ym3，对Pb和Zn吸收量最大的分别为ym1和ym2；种子对Pb和Cd吸收量最大的为ym2，对As和Cu吸收量最大的为ym5，对Zn吸收量最大则为ym3，ym1种子对Pb、As和Cu吸收量皆最小，对Cd、Zn吸收最小的品种分别为ym4和ym2；5个品种表现为除Cu外，茎叶富集系数皆大于根，根系向茎叶的转运系数均大于1。研究表明，ym1可作为Pb污染修复植物，ym3可作为As、Zn、Cu和Cd污染修复的适推品种。     

重金属污染土壤几种生物修复方式比较

土壤重金属污染是全球普遍存在的问题,生物修复因其环境友好且成本效益高而得到广泛关注。但不同生物修复技术有其优势和局限性,充分了解每种修复技术的特点,才能更经济、有效地对污染土壤进行修复。本研究阐述对比了目前的土壤重金属生物修复方法,包括植物修复（植物挥发、植物固定和植物提取）、转基因植物提取、螯合辅助植物修复、微生物辅助植物修复等技术的机制、优势、局限性和适用性等方面的差异。综述提出有效的生物修复技术需要土壤化学、植物生物学、遗传学、微生物学和环境工程等多学科的有机结合。根据污染土壤的特点,结合具有相应改良特性的转基因植物,是实现污染土壤大面积修复的有效方法。同时,农艺措施对天然超级积累植物的生物量和重金属提取能力的刺激作用还需要进一步挖掘。植物修复可以与其他几种传统修复技术有效结合,利用转基因技术建立土壤+植物+微生物的组合是未来修复技术发展的最佳途径。     

根瘤菌在污染土壤修复中的地位和作用

根瘤菌是一种在环境中广泛存在,常与豆科植物共生结瘤固氮的细菌类群,在农、林、牧业的可持续发展中具有重要作用。近年来研究发现,除了人们熟知的固氮功能,根瘤菌还参与环境中重金属、有机物等污染土壤的修复,在生态环境修复、维持土地生产力以及节能减排方面发挥了重要的作用,尤其是根瘤菌与豆科植物联合修复污染环境效果更为显著。本文综合介绍了根瘤菌在重金属、有机物等污染土壤及滨海盐渍土壤修复中的地位、作用及潜在价值,以期为改善生态环境提供一些可参考的方法和依据。     

降解菌LW3对氯嘧磺隆污染土壤的生物修复研究

为了探讨添加外源接种细菌对土壤氯嘧磺隆污染修复的影响，利用从长期污染土壤分离到的氯嘧磺隆降解菌LW3，在实验室模拟情况下，研究其对氯嘧磺隆污染土壤的修复情况及其影响因素。结果表明，降解菌接种量越高，氯嘧磺隆降解效果越好，但在接种量到达1.0×10⁶ CFU/g干土后，再增加接种量对降解率的增加效果就不太明显。土壤降解的最适温度为30℃，最适初始pH 7。在土壤含水量低于40%时，菌株对土壤氯嘧磺隆的降解率随着土壤的含水量的升高而增加；而在淹水情况下时，氯嘧磺隆的降解受到严重抑制。

     

镉胁迫对尾巨桉幼苗生长和生理特性的影响

以尾巨桉(DH_32-29)幼苗为试验材料，以植株的生长指标(苗高、地径、叶面积)、生物量、根冠比、氮平衡指数(NBI)、叶绿素相对含量(SPAD 值)、丙二醛(MDA)含量和叶绿素荧光参数为指标，通过盆栽试验设置8个10~140 mg·kg^-1浓度镉(Cd)盐溶液处理，研究了不同浓度 Cd 胁迫处理下尾巨桉幼苗的形态和生理特性变化规律。结果显示：低浓度(10 mg·kg^-1)Cd 处理对尾巨桉幼苗的形态和生理特性无明显影响。当 Cd 浓度大于20 mg·kg^-1，随着 Cd浓度的逐渐增大，其对尾巨桉幼苗的伤害程度也逐渐增加，主要表现为：植株生长缓慢、生物量减小、叶绿素相对含量和叶绿素荧光参数(Fv/Fm、φPSⅡ、NP、NBI)均下降，qP 无明显变化， MDA 含量升高。Cd 浓度大于60 mg·kg^-1以上时，尾巨桉幼苗生长出现一定程度上的损伤症状和生长停滞现象。结果表明低浓度(≤20 mg·kg^-1)Cd处理下尾巨桉幼苗能正常生长，随着浓度的增大，Cd 处理对尾巨桉的形态和生理特性造成显著影响。同时结合部分桉树适生区土壤 Cd 含量调查及相关研究结果认为，尾巨桉具有一定修复 Cd 污染区土壤的潜力。